TIME-DEPENDENT miRNA PROFILE AND RENAL ACUTE PHASE RESPONSE DURING SEPTIC ACUTE KIDNEY INJURY IN MICE
Róka Beáta
Elméleti és Transzlációs Orvostudományok
Dr. Kellermayer Miklós
SE, Elméleti Orvostudományi Centrum, Hevesy György előadóterem
2024-04-25 14:30:00
A folyadék- és elektrolitháztartás szabályozásának élet- és kórélettana
Dr. Zsembery Ákos
Dr.Hamar Péter Dr.Szénási Gábo
Dr. Hosszú Ádám
Dr. Nagy Béla
Dr. Reusz György
Dr. Kuthi Levente
Dr. Remport Ádám
Bacterial lipopolysaccharide-induced systemic inflammation and circulatory shock is associated with acute kidney injury (AKI). We investigated the temporal changes in renal miRNA and protein expression induced by endotoxin shock in mice.
AKI was induced in male outbred NMRI mice by intraperitoneal LPS injections. Mice were sacrificed at 1.5 and 6 h (early phase, EP) or at 24 and 48 h (late phase, LP) after LPS injection. The temporal miRNome profile of septic AKI was established using miRCURY LNA™ miRNA microarray and verified by qPCR. Total renal proteome was analysed by HPLC-MS/MS.
AKI was confirmed by increased plasma urea concentrations and enhanced renal mRNA expression of IL-6, Tnf-α and Lcn-2. Renal miRNome and proteome changes were mild at 1.5 h. We detected the strongest miRNA dysregulation at 6 and 24 h that started to normalize by 48 h. In EP, miR-762 was the most upregulated miRNA that we newly associated with septic AKI. A predicted target of miR-762, Ras related GTPase 1B (Sar1b) was downregulated, indicating that miR-762 may regulate GTP signalling. In LP, miR-21a-5p was the most induced miRNA followed by miR-144-3p, miR-146a-5p and miR-451a. Renal protein concentration of potential protein targets of miR-144-3p was downregulated in LP, which were: aquaporin-1 (Aqp1), solute carrier family 22 member 8 (Slc22a8 or Oat3) and electrogenic sodium bicarbonate cotransporter 1 (Slc4a4 or NBCe1). These miRNAs and transporters may play an important role in LPS-induced AKI and renal inflammation.
As part of the renal inflammation induced by LPS, we demonstrated a massive, complex and coordinated local acute-phase response (APR) of the kidney. APPs dominated the proteome in LP (the ratio of APPs among proteins upregulated at least 4-fold was: EP, 1.5 h: 0/10, 6 h: 1/10; LP, 24 h: 22/47, 48 h: 17/44). Lipocalin-2, complement C3, fibrinogen, haptoglobin and hemopexin were the most upregulated APPs. Renal mRNA expression of APPs was measured by qPCR. The mRNA expression of APPs was induced in EP, preceding the changes in protein concentrations, and confirming local renal production of most APPs.
